- Proteome and metabolome analyses reveal differential responses in tomato -Verticillium dahliae-interactions. [Journal Article]
- JPJ Proteomics 2019 Jul 16; :103449
- Verticillium dahliae colonizes vascular tissue and causes vascular discoloration in susceptible hosts. Two well-defined races exist in V. dahliae populations from tomato and lettuce. In this study, p…
Verticillium dahliae colonizes vascular tissue and causes vascular discoloration in susceptible hosts. Two well-defined races exist in V. dahliae populations from tomato and lettuce. In this study, proteins and metabolites obtained from stems of race 1-incompatible (Beefsteak) and -compatible (Early Pak) tomato cultivars were characterized. A total of 814 and 584 proteins in Beefsteak; and 456 and 637 proteins in Early Pak were identified in stem extracts of plants inoculated with races 1 and 2, respectively. A significant number of defense-related proteins were expressed in each tomato-V. dahliae interaction, as anticipated. However, phenylalanine ammonia-lyase (PAL), an important defense-associated enzyme of the phenylpropanoid pathway, in addition to remorin 1, NAD-dependent epimerase/dehydratase, and polyphenol oxidase were uniquely expressed in the incompatible interaction. Compared with the uninoculated control, significant overexpression of gene ontology terms associated with lignin biosynthesis, phenylpropanoid pathway and carbohydrate methylation were identified exclusively in the incompatible interaction. Phenolic compounds known to be involved in plant defense mechanisms were at higher levels in the incompatible relative to the compatible interactions. Based on our findings, PAL and enzymes involved defense-related secondary metabolism and the strengthening of cell walls is likely critical to confer resistance to race 1 of V. dahliae in tomato. SIGNIFICANCE: Verticillium dahliae, a soilborne fungal pathogen and a widely distributed fungal pathogen, colonizes vascular tissue and causes vascular discoloration in roots and stems, leaf wilting, and death of susceptible plant hosts. It causes billions of dollars in annual crop losses all over the world. The study focused on the proteomic and metabalomic of V. dahliae interactions (incompatible with Beefsteak and compatible with Early Pak tomato cultivars). Based on our findings, PAL and enzymes involved defense-related secondary metabolism and the strengthening of cell walls is likely critical to confer resistance to race 1 of V. dahliae in tomato.
- A potential probiotic Chromobacterium aquaticum with bacteriocin-like activity enhances the expression of indicator genes associated with nutrient metabolism, growth performance and innate immunity against pathogen infections in zebrafish (Danio rerio). [Journal Article]
- FSFish Shellfish Immunol 2019 Jul 16
- The use of probiotics as alternatives to antibiotics for disease control is a relatively eco-friendly approach in aquaculture; hence, studies isolating and assessing the benefit of potential probioti…
The use of probiotics as alternatives to antibiotics for disease control is a relatively eco-friendly approach in aquaculture; hence, studies isolating and assessing the benefit of potential probiotics to fish farming are common. The zebrafish is an excellent model system for validating beneficial functions of potential probiotics before their practical application in aquaculture. Here, a potentially probiotic Chromobacterium aquaticum was isolated from lake water samples and characterized by biochemical analysis and 16S rDNA sequencing. The probiotic produced extracellular enzymes (protease and xylanase) and a bacteriocin-like substance, which exhibited tolerance to extreme pH and high-temperature conditions and broad-spectrum bactericidal activity against diverse pathogens, including aquatic, foodborne, clinical and plant pathogens. The effects of C. aquaticum on zebrafish nutrient metabolism, growth performance and innate immunity were evaluated by measuring the expression of indicator genes after C. aquaticum feeding for 8 weeks. Fish administered the probiotic exhibited significantly increased hepatic mRNA expression of carbohydrate metabolism-related genes, including glucokinase (GK), hexokinase (HK), glucose-6-phosphatase (G6Pase), and pyruvate kinase (PK-L), and growth-related genes, including the growth hormone receptor (GHR) and insulin-like growth factor-1 (IGF-1). Innate immune-related genes (IL-1β, IL-6, TNF-α, IL-10, IL-21, NF-κb, lysozyme and complement C3b) were induced in fish with probiotic supplementation. Probiotic-treated fish exhibited a higher survival rate than control fish after challenge with Aeromonas hydrophila and Streptococcus iniae. Together, these data suggest that C. aquaticum, as a probiotic feed supplement, could enhance nutrient metabolism and growth performance and could modulate innate immunity against A. hydrophila and S. iniae in zebrafish.
- Acute phase protein response to viral infection and vaccination. [Review]
- ABArch Biochem Biophys 2019 Jul 16
- Organisms respond in multiple ways to microbial infections. Pathogen invasion tipically triggers an inflammatory response where acute phase proteins (APP) have a key role. Pentraxins (PTX) are a fami…
Organisms respond in multiple ways to microbial infections. Pathogen invasion tipically triggers an inflammatory response where acute phase proteins (APP) have a key role. Pentraxins (PTX) are a family of highly conserved APP that play a part in the host defense against infection. The larger proteins of the family are simply named pentraxins, while c-reactive proteins (CRP) and serum amyloid proteins (SAA, SAP) are known as short pentraxins. Although high APP levels have been broadly associated with bacterial infections, there is a growing body of evidence revealing increased PTX, CRP and SAP expression upon viral infection. Furthermore, CRP, PTX and SAP have shown their potential as diagnostic markers and predictors of disease outcome. Likewise, the measurement of APP levels can be valuable to determine the efficacy of antiviral therapies and vaccines. From the practical point of view, the ability of APP to reduce viral infectivity has been observed in several virus-host models. This has prompted investigation efforts to assess the role of acute phase response proteins as immunoregulatory molecules and their potential as therapeutic reagents. This work aims to present an overview of the APP response to viral infections reviewing the current knowledge in the field.
- Peer pressure from a Proteus mirabilis self-recognition system controls participation in cooperative swarm motility. [Journal Article]
- PPPLoS Pathog 2019 Jul 19; 15(7):e1007885
- Colonies of the opportunistic pathogen Proteus mirabilis can distinguish self from non-self: in swarming colonies of two different strains, one strain excludes the other from the expanding colony edg…
Colonies of the opportunistic pathogen Proteus mirabilis can distinguish self from non-self: in swarming colonies of two different strains, one strain excludes the other from the expanding colony edge. Predominant models characterize bacterial kin discrimination as immediate antagonism towards non-kin cells, typically through delivery of toxin effector molecules from one cell into its neighbor. Upon effector delivery, receiving cells must either neutralize it by presenting a cognate anti-toxin as would a clonal sibling, or suffer cell death or irreversible growth inhibition as would a non-kin cell. Here we expand this paradigm to explain the non-lethal Ids self-recognition system, which stops access to a social behavior in P. mirabilis by selectively and transiently inducing non-self cells into a growth-arrested lifestyle incompatible with cooperative swarming. This state is characterized by reduced expression of genes associated with protein synthesis, virulence, and motility, and also causes non-self cells to tolerate previously lethal concentrations of antibiotics. We show that temporary activation of the stringent response is necessary for entry into this state, ultimately resulting in the iterative exclusion of non-self cells as a swarm colony migrates outwards. These data clarify the intricate connection between non-lethal recognition and the lifecycle of P. mirabilis swarm colonies.
- Sources of Inoculum and Survival of Macrophomina phaseolina in Florida Strawberry Fields. [Journal Article]
- PDPlant Dis 2019 Jul 19; :PDIS03190510RE
- Macrophomina phaseolina, the causal agent of charcoal rot, affects strawberry crowns, inducing plant collapse. The fungus survives in the soil through the production of microsclerotia and is usually …
Macrophomina phaseolina, the causal agent of charcoal rot, affects strawberry crowns, inducing plant collapse. The fungus survives in the soil through the production of microsclerotia and is usually controlled by preplant fumigation of soil. However, in the 2016 to 2017 Florida strawberry season, even after soil fumigation, about 30% plant mortality still occurred in plastic-covered beds that were used for a second season and where crop residue (mainly old strawberry crowns) was disposed of between beds. Therefore, this study was conducted to determine if M. phaseolina can survive on strawberry debris over summer in Florida and if so, verify whether strawberry debris might act as a source of inoculum for new transplants. Crowns from the previous season were collected from commercial farms where charcoal rot had been reported, and M. phaseolina was recovered from all samples. In a research field, infected crowns were buried in the soil at different depths and retrieved every 2 weeks during the summer. After 8 weeks, M. phaseolina could be recovered at all depths. Moreover, inoculation of strawberry plants by drenching the soil, dipping roots, or spraying leaves with a M. phaseolina microsclerotial suspension from pure cultures or infected crowns produced symptoms with differences in incubation periods depending on cultivar susceptibility. Furthermore, infected crowns disposed of in the aisles between beds or buried next to new transplants of cultivars Strawberry Festival, Florida Beauty, and Winterstar induced charcoal rot, with the level of aggressiveness depending on the cultivar susceptibility and inoculum placement in the field.
- What Is the Role of Molecular Techniques for Detection of Pathogen Deoxyribonucleic Acid (DNA) (Polymerase Chain Reaction [PCR] or Next-Generation Sequencing) in Patients With Infected Total Ankle Arthroplasty (TAA)? [Journal Article]
- FAFoot Ankle Int 2019; 40(1_suppl):29S-31S
- Molecular techniques, particularly next-generation sequencing and the Ibis T5000 technology, have the potential to be used as an important adjunct in the diagnosis of bacterial infection following to…
Molecular techniques, particularly next-generation sequencing and the Ibis T5000 technology, have the potential to be used as an important adjunct in the diagnosis of bacterial infection following total ankle arthroplasty (TAA), although sufficient clinical evidence is lacking.
- Is There a Role for Measuring Synovial Biomarkers for Diagnosis of Infected Total Ankle Arthroplasty (TAA)? [Journal Article]
- FAFoot Ankle Int 2019; 40(1_suppl):27S-29S
- Based on the hip and knee arthroplasty literature, measuring synovial biomarkers may play a role in the diagnosis of infected total ankle arthroplasty (TAA). The diagnosis of periprosthetic joint inf…
Based on the hip and knee arthroplasty literature, measuring synovial biomarkers may play a role in the diagnosis of infected total ankle arthroplasty (TAA). The diagnosis of periprosthetic joint infection (PJI) in the setting of a TAA can be confirmed with cultures, provided that a plausible pathogen is recovered in the context of a compatible clinical picture. In the absence of a positive culture, synovial biomarker analysis may help in establishing the diagnosis.
- Strategies to Counteract Botulinum Neurotoxin A: Nature's Deadliest Biomolecule. [Journal Article]
- ACAcc Chem Res 2019 Jul 19
- Botulinum neurotoxin serotype A (BoNT/A), marketed commercially as Botox, is the most toxic substance known to man with an estimated intravenous lethal dose (LD50) of 1-2 ng/kg in humans. Despite its…
Botulinum neurotoxin serotype A (BoNT/A), marketed commercially as Botox, is the most toxic substance known to man with an estimated intravenous lethal dose (LD50) of 1-2 ng/kg in humans. Despite its widespread use in cosmetic and medicinal applications, no postexposure therapeutics are available for the reversal of intoxication in the event of medical malpractice or bioterrorism. Accordingly, the Centers for Disease Control and Prevention categorizes BoNT/A as a Category A pathogen, posing the highest risk to national security and public health as a result of the ease with which BoNT/A can be weaponized and disseminated. BoNT/A-mediated lethality results from neurons impeded from releasing acetylcholine, which ultimately causes muscle paralysis and possible death by asphyxiation with the loss of diaphragm function. Currently, the only available respite for BoNT/A poisoning is antibody-based therapy; however, this intervention is only effective within 12-24 h postexposure. Small molecule therapeutics remain the only opportunity to reverse BoNT/A intoxication after neuronal poisoning and are urgently needed. Nevertheless, no small molecule BoNT/A inhibitors have reached the clinic or even advanced to clinical trials. This Account highlights the accomplishments and existing challenges facing BoNT/A drug discovery today. Using the comprehensive body of work from our laboratory, we illustrate our nearly two-decade endeavor to discover a clinically relevant BoNT/A inhibitor. Specifically, a discussion on the identification and characterization of new chemical leads, the development of in vitro and in vivo assays, and pertinent discoveries in BoNT/A structural biology related to small molecule inhibition is presented. Lead discovery efforts in our laboratory have leveraged both in vitro high-throughput screening and rational design, and an array of mechanistic strategies for inhibiting BoNT/A has been discovered, including noncovalent inhibition, metal-binding active site inhibition, covalent inhibition, and α- and β-exosite inhibition. We contrast the strengths and weaknesses of each of these mechanistic strategies and propose the most favorable approach for success. Finally, we discuss multiple serendipitous discoveries of antibotulism small molecules with alternative mechanisms of action. Remaining challenges facing clinically relevant BoNT/A inhibition are presented and analyzed, including the current inability to reconcile toxin half-life (months to greater than one year) in neurons with in vivo pharmaceutical lifetimes and reoccurring inconsistencies between in vitro, cellular, and in vivo translation. Our Account of BoNT/A chemical research emphasizes the present accomplishments and critically analyzes the remaining obstacles for drug discovery. Importantly, we call for an increased focus on the discovery of safe and effective covalent inhibitors of BoNT/A that compete with the inherent half-life of the toxin.
- ULTRASOUND DIAGNOSTICS OF CERVICAL CHANGES WITH DIFFERENT LOCALIZATION OF THE CERVICITIS IN WOMEN WITH THE ECTOPY OF THE VAGINAL PORTION OF THE CERVIX. [Journal Article]
- GMGeorgian Med News 2019; (290):32-37
- The aim of the study was to improve the diagnosis of structural changes in all parts of the cervix depending on the preferential localization of the inflammatory process - in the ectocervix, endocerv…
The aim of the study was to improve the diagnosis of structural changes in all parts of the cervix depending on the preferential localization of the inflammatory process - in the ectocervix, endocervix, and also in the stroma in women with ectopia (pseudoerosis) of the vaginal part of the cervix. The study included women of predominantly early and middle reproductive age, as these women have inflammatory processes more often than in menopause. To perform the work, a transvaginal imaging method was used, which allowed us to see the smallest structures (up to 0.5-1.0 mm) of the cervix. We have previously developed a technique for visualizing the external part of the cervix and obtained a patent of Ukraine. The results of the ultrasound were compared with laboratory data, colposcopy and cervicoscopy. Traditionally, the diagnosis of cervicitis was made on the basis of clinical symptoms, smear results to determine the presence and type of pathogen. Endoscopic methods allowed visualization of only the mucous membrane of the cervical canal and external part of the cervix. The deep layers of the ectocervix, endocervix and cervical stroma can only be visualized using high-frequency ultrasound in a transvaginal way. According to the results of the study, it was possible to determine the preferential localization of the inflammatory process, to determine the nature of changes in different layers of the cervix. These changes were expressed in the presence of calcifications and cystic cavities of various sizes (1-6 mm), an increase or decrease in echogenicity, the degree of heterogeneity of the structure of the ecto-, endocervix and stroma.
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- Inheritance of Virulence and Linkages of Virulence Genes in an Ethiopian Isolate of the Wheat Stripe Rust Pathogen (Puccinia striiformis f. sp. tritici) Determined Through Sexual Recombination on Berberis holstii. [Journal Article]
- PDPlant Dis 2019 Jul 19; :PDIS02190269RE
- Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most devastating wheat diseases in Ethiopia. To study virulence genetics of the pathogen, 117 progeny isolates were produced …
Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most devastating wheat diseases in Ethiopia. To study virulence genetics of the pathogen, 117 progeny isolates were produced through sexual reproduction of an Ethiopian isolate of the stripe rust pathogen on Berberis holstii plants under controlled conditions. The parental and progeny isolates were characterized by phenotyping on wheat lines carrying single Yr genes for resistance and genotyped using 10 polymorphic simple sequence repeated (SSR) markers. The progeny isolates were classified into 37 virulence phenotypes and 75 multilocus genotypes. The parental isolate and progeny isolates were all avirulent to resistance genes Yr5, Yr10, Yr15, Yr24, Yr32, YrTr1, YrSP, and Yr76 but virulent to Yr1 and Yr2, indicating that the parental isolate was homozygous avirulent or homozygous virulent at these loci. The progeny isolates segregated for virulence to 12 Yr genes. Virulence phenotypes to Yr6, Yr28, Yr43, and Yr44 were controlled by a single dominant gene; those to Yr7, Yr9, Yr17, Yr27, Yr25, Yr31, and YrExp2 were each controlled by two dominant genes; and the virulence phenotype to Yr8 was controlled by two complementary dominant genes. A linkage map was constructed with seven SSR markers, and 16 virulence loci corresponding to 11 Yr resistance genes were mapped with some loci linked to each other. These results are useful in understanding host-pathogen interactions and selecting resistance genes to develop wheat cultivars with highly effective resistance to stripe rust.